Journal: PLoS ONE

Article Title: Comparative Anterior Pituitary miRNA and mRNA Expression Profiles of Bama Minipigs and Landrace Pigs Reveal Potential Molecular Network Involved in Animal Postnatal Growth

doi: 10.1371/journal.pone.0131987

Figure Lengend Snippet: Differentially expressed miRNAs between Bama minipigs (YB) and Landrace (YC).

Article Snippet: Six hours after transfection, the second transfection were performed with their corresponding miRNAs mimics (ssc-miR-7139-3p for FSHB, ssc-miR-133a-3p for GNAI3) or a negative control (10pmol/well, GenePharma, Shanghai, China).

Techniques:

Journal: PLoS ONE

Article Title: Comparative Anterior Pituitary miRNA and mRNA Expression Profiles of Bama Minipigs and Landrace Pigs Reveal Potential Molecular Network Involved in Animal Postnatal Growth

doi: 10.1371/journal.pone.0131987

Figure Lengend Snippet: (A) The second structure of Y-47(custom-designed miRNA) and its potential target FSHB (for seed sequence only). (B) The second structure of ssc-miR-133a-3p and its potential target GNAI3 (for seed sequence only). (C)Y-47 mimics significantly decreased FSHB construct luciferase activity (relative, firefly luciferase activity/Renilla luciferase activity). (D) ssc-miR-133a-3p mimics significantly decreased GNAI3 construct luciferase activity (relative, firefly luciferase activity/Renilla luciferase activity). Statistical significance was determined by ANOVA, followed by Tukey’s multiple comparisons test, the panels with different letter were considered statistically significant (p<0.05).

Article Snippet: Six hours after transfection, the second transfection were performed with their corresponding miRNAs mimics (ssc-miR-7139-3p for FSHB, ssc-miR-133a-3p for GNAI3) or a negative control (10pmol/well, GenePharma, Shanghai, China).

Techniques: Sequencing, Construct, Luciferase, Activity Assay

Journal: Frontiers in Physiology

Article Title: Two Putative Cypovirus-Encoded miRNAs Co-regulate the Host Gene of GTP-Binding Nuclear Protein Ran and Facilitate Virus Replication

doi: 10.3389/fphys.2021.663482

Figure Lengend Snippet: Regulation of BmCPV-miRNAs on the target gene in HEK293T cells and in BmN cells. (A) BmCPV-miRNAs were expressed in the transfected HEK293T cells. (B) BmCPV-miRNAs downregulate the expression of the substituent target gene mCherry in HEK293T cells. pLNHX-Ran: pLNHX-mCherry-RanUTR; pLNHX-Ran-mut: pLNHX-mCherry-RanUTR-mut. (C) Regulation of BmCPV-miRNA mimics on expression of target gene BmRan in BmN cells ( n = 3, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: To verify the miRNA regulation on target gene expression in BmN cells, miRNA mimics were transfected into the BmN cells to detect the changes in expression of the target gene. miRNA mimics and NC (sequences shown in ) were synthesized by Shanghai GenePharma Co., Ltd. BmN cells were inoculated into a six-well plate 16 h before transfection in which the cell density should be over 60%, and they were divided into three groups, i.e., blank control, NC, and mimics, each group was set for three replicates.

Techniques: Transfection, Expressing

Journal: Journal of Cancer

Article Title: Brown Seaweed Fucoidan Inhibits Cancer Progression by Dual Regulation of mir-29c/ADAM12 and miR-17-5p/PTEN Axes in Human Breast Cancer Cells

doi: 10.7150/jca.15703

Figure Lengend Snippet: Fucoidan modulates microRNAs to target the 3'-UTR region of ADAM12 and PTEN and represses their expression in human breast cancer cells. (A) (C) Schematic representation of 3'-UTRs of ADAM12 and PTEN showing the putative miR-29c and miR-17-5p target sites. (B) (D) After transfection of cells, the ADAM12- and PTEN-dependent luciferase activity of breast cancer cells was measured using the Dual-Luciferase ® Reporter Assay System 48 h after treatment, as indicated. The experiments were performed in triplicate. Values were normalized to Renilla luciferase activity (transfection efficiency control) and are plotted as relative luciferase activities. SCR, scrambled control. Significant differences are shown (*p < 0.05 and **p < 0.01, compared with the control group).

Article Snippet: The miR-29c, miR-17-5p, or scrambled miRNA mimic (100 nM); ADAM12 and PTEN 3'-UTR firefly luciferase report vectors (wild-type or mutated type); and the control vector containing the Renilla luciferase pRL-TK vector (Promega) were cotransfected into breast cancer cells by using Lipofectamine-2000 transfection reagent (Invitrogen), according to the manufacturer's protocol.

Techniques: Expressing, Transfection, Luciferase, Activity Assay, Reporter Assay, Control